174 THE VITAMINES 



purification; it was dissolved in water and precipitated with a 

 20 per cent tannin solution till only a slight cloudiness was seen. 

 The precipitate was filtered off and quickly washed with a 1 

 per cent tannin solution. It was then rubbed up in a mortar 

 with 3 per cent sulphuric acid till a complete solution resulted. 

 This was treated with an excess of baryta, the precipitate filtered 

 off, and the excess of baryta removed from the filtrate with 

 dilute sulphuric acid. The solution was shaken out with ether and 

 concentrated. In this way, a preparation was obtained, called crude 

 oryzanin II., which was three times as active as the first preparation. 

 When a concentrated watery solution of this preparation was 

 rubbed up with a slight excess of dilute picric acid, a flocculent 

 precipitate settled out which became crystalline on standing in the 

 cold. These crystals still occluded some nicotinic acid picrate, but 

 with careful technique the latter remained in solution. The oryzanin 

 picrate was recrystallized by dissolving in cold acetone and allowing 

 this to evaporate in the dessicator; yellowish brown microscopic 

 needles, grouped in star formation, were obtained. An amount 

 corresponding to two centigrams of picrate was very active for 

 pigeons; the substance was given only to two pigeons. The amount 

 of picrate obtained was so slight that there was not enough for a 

 melting point. The question as to whether the pure oryzanin would 

 give the same decomposition products as oryzanin I. was therefore 

 left open by the authors. Since the publication of this work in 

 1912, nothing of a corroborative nature has been printed by the 

 Japanese investigators (at least to our knowledge). In the mean- 

 time, Drummond and Funk (481) tried to confirm the above findings, 

 but all attempts to isolate the substance as a picrate failed. Above 

 all, it was evident that an extract of rice polishings is still quite a 

 complicated mixture. Of the many substances it contains, we were 

 able to isolate, besides the previously mentioned choline, allantoine 

 and nicotinic acid, also betaine, adenine, guanine and apparently 

 guanidine. An observation, made accidently, showed us how careful 

 one must be in drawing conclusions from such fractionations. In this 

 instance, an apparently pure substance was isolated, which had a 

 constant melting point on recrystallization. The substance consisted 

 of betaine and nicotinic acid, which could not be separated from each 

 other by recrystallization. Only when the nicotinic acid was 

 separated as a copper salt, was the betaine apparent. It was also 



